LOCUS       sequence-441689-        4767 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor plasmid of hAT-14_Crp Transposon.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4767)
  AUTHORS   Zhang T, Tan S, Tang N, Li Y, Zhang C, Sun J, Guo Y, Gao H, Cai Y, 
            Sun W, Wang C, Fu L, Ma H, Wu Y, Hu X, Zhang X, Gee P, Yan W, Zhao 
            Y, Chen Q, Guo B, Wang H, Zhang YE
  TITLE     Heterologous survey of 130 DNA transposons in human cells highlights
            their functional divergence and expands the genome engineering 
            toolbox.
  JOURNAL   Cell. 2024 Jul 11;187(14):3741-3760.e30. doi: 
            10.1016/j.cell.2024.05.007. Epub 2024 Jun 5.
  PUBMED    38843831
REFERENCE   2  (bases 1 to 4767)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.cell.2024.05.007"; journalName: "Cell"; date: 
            "2024-07-11- 11"; volume: "187"; issue: "14"; pages: "3741-3760.e30"
COMMENT     Sequence Label: pMV-hAT-14_Crp
FEATURES             Location/Qualifiers
     source          1..4767
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     94..112
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     121..138
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     122..138
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        409..908
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(444..467)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     841..860
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      922..931
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             928..1527
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MGTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIE
                     RVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRL
                     AAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLET
                     SAPRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     1427..1447
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             1534..1590
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             join(1591..1593,1594..1596,1597..2310)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1591..1593,1594..1596,1597..2307)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1636..1657)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1897..1916)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2244..2265
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2332..2349)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2338..2562
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2870..2886)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2870..2886)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2898..2915)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3069..3657)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3149..3168)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3828..4619,4620..4688))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLRNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERKRQIAEIGAS
                     LIKHW"
     primer_bind     4451..4470
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(4689..4767,1..26))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
       61 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
      121 tgtaaaacga cggccagtcg aaccacgcaa tgcgtctcga tccgcagtgt cttgcgtctc
      181 tctgcagcta tatagtaggt taaagatgga gcggtcgaaa agaagccatg tgtggaatca
      241 tttcacaaaa atcaaagaag atgctgttag atgtaagatg tgcaatacag agcttaggta
      301 cctgaacagc actggagcaa tgctgaatca tcttaagctg aagcatccct ctgtttcaag
      361 cgaaagacaa gatcagaaac aaacaacacc ggtgctagca attctaccgg gtaggggagg
      421 cgcttttccc aaggcagtct ggagcatgcg ctttagcagc cccgctgggc acttggcgct
      481 acacaagtgg cctctggcct cgcacacatt ccacatccac cggtaggcgc caaccggctc
      541 cgttctttgg tggccccttc gcgccacctt ctactcctcc cctagtcagg aagttccccc
      601 ccgccccgca gctcgcgtcg tgcaggacgt gacaaatgga agtagcacgt ctcactagtc
      661 tcgtgcagat ggacagcacc gctgagcaat ggaagcgggt aggcctttgg ggcagcggcc
      721 aatagcagct ttgctccttc gctttctggg ctcagaggct gggaaggggt gggtccgggg
      781 gcgggctcag gggcgggctc aggggcgggg cgggcgcccg aaggtcctcc ggaggcccgg
      841 cattctgcac gcttcaaaag cgcacgtctg ccgcgctgtt ctcctcttcc tcatctccgg
      901 gcctttcgac ctgcagccaa cgccaccatg gggaccgagt acaagcccac ggtgcgcctc
      961 gccacccgcg acgacgtccc ccgggccgta cgcaccctcg ccgccgcgtt cgccgactac
     1021 cccgccacgc gccacaccgt cgacccggac cgccacatcg agcgggtcac cgagctgcaa
     1081 gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc
     1141 gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag
     1201 atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggaa
     1261 ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc
     1321 tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcg
     1381 gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc
     1441 tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc
     1501 tggtgcatga cccgcaagcc cggtgccatc gatgccacta acttctccct gttgaaacaa
     1561 gcaggggatg tcgaagagaa tcccgggcca atggtgagca agggcgagga gctgttcacc
     1621 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
     1681 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc
     1741 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag
     1801 tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
     1861 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc
     1921 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
     1981 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac
     2041 gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac
     2101 aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
     2161 gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa
     2221 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
     2281 actctcggca tggacgagct gtacaagtaa ttaattaagc tcgctgatca gcctcgactg
     2341 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
     2401 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
     2461 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
     2521 aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag gcgcgccctc
     2581 gagaaatcat caatagagtt gtgatgcagc tttttgtttt tgttgttgtt gcttgtttaa
     2641 cctcaacaga accttgagat gtgaggttgt tttttttttt ctgattggat tttttttttc
     2701 atttttggaa gacaaaacat tttttaatga ggattttatg taaaaaaaaa aattgttgga
     2761 gtgtattgtt cttgaagttt taactggatc cttttgcact tttggaaggc aaaacaagga
     2821 tttaatacta tataggggcc cagagacgga gtcactgcca accgagacgg tcatagctgt
     2881 ttcctgtgtg ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     2941 gcggtatcag ctcactcaaa ggcggtaata cggttaccca cagaatcagg ggataacgca
     3001 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     3061 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     3121 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     3181 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     3241 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     3301 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     3361 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     3421 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     3481 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     3541 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     3601 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     3661 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     3721 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     3781 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     3841 atcagtgagg cacctatctc agcgatctgt ctctttcgtt catccatagt tgcctgactc
     3901 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaata
     3961 ataccgcggg acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     4021 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     4081 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatc
     4141 gctacaggca tcgtggtatc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     4201 caacgatcaa ggcgagttac atgatccccc atgttgcgca aaaaagcggt tagctccttc
     4261 ggtcctccga tcgttgtcag aagtaagttg gccgccgtgt tatcactcat ggttatggca
     4321 gcactacata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     4381 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     4441 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     4501 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     4561 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     4621 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     4681 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     4741 agcggataca tatttgaatg tatttag
//
